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目的探讨人类免疫缺陷病毒/艾滋病(HIV/AIDS)患者中和抗体保守表位氨基酸变异特征,为开展中和抗体免疫治疗和疫苗设计奠定理论基础。方法RTPCR及nestPCR扩增HIV/AIDS患者HIV1外膜envC2～C3区基因,双脱氧终止法进行核酸序列测定,翻译为氨基酸序列,并与HIV1SequenceDatabase参考毒株中和抗体表位数据比对辨别其保守表位氨基酸突变情况。结果HIV1外膜蛋白gp120C2～C3区,HIV感染者和AIDS患者CD4结合位点(CD4BS)、CD4诱导(CD4i)、2G12三种类型中和抗体保守表位氨基酸均存在突变,两组病例表位突变率差异无统计学意义(均P>0.05);CD4BS保守表位突变有370E/Q(12.1%),370E/K(4.5%),266A/S(1.5%),368D/E(1.5%);CD4i保守表位突变有370E/Q(12.1%),370E/K(4.5%),381E/D(7.6%),381E/V(1.5%);2G12保守表位突变有295N/V(19.7%),295N/T(6.1%),295N/D、295N/K、295N/E(各1.5%),297T/I(9.1%),297T/S、297T/N(各1.5%)。结论我国HIV/AIDS患者HIV1外膜蛋白gp120C2～C3区中和抗体CD4BS、CD4i、2G12保守表位氨基酸均存在突变,但尚处于低突变水平;不同类型中和抗体保守表位各氨基酸位点的变异程度有差异。
Objective To investigate the characteristics of amino acid variations in the conserved epitopes of neutralizing antibodies in patients with human immunodeficiency virus / AIDS (HIV / AIDS) and to lay a theoretical foundation for the immunotherapy and vaccine design of neutralizing antibodies. Methods RTPCR and nestPCR were used to amplify env C2 ~ C3 gene of HIV1 from HIV / AIDS patients. The nucleic acid sequences were determined by dideoxy terminator method and translated into amino acid sequence. The amino acid sequence was compared with the HIV1 Sequence Database neutralizing antibody epitope data to identify the conservative Epitope amino acid mutations. Results There was a mutation in the conserved epitope amino acids of CD4 +, CD4i and 2G12 neutralizing antibodies in the gp120C2 ~ C3 region of HIV1 outer membrane protein, HIV infection and AIDS patients. The epitopes of two groups There were no significant differences in the mutation rates between the two groups (all P> 0.05). The conservative frequency of CD4BS mutation was 370E / Q (12.1%), 370E / K ). The conserved epitopes of CD4i were 370E / Q (12.1%), 370E / K (4.5%), 381E / D 297 T / I (29.7%), 295 N / T (6.1%), 295 N / D, 295 N / K, 295 N / E (1.5% each), 297 T / I (9.1%), 297 T / S, 297 T / N (1.5% each). CONCLUSIONS: The conservative amino acid residues of CD4BS, CD4i and 2G12 of HIV1 outer membrane protein gp120C2 ~ C3 in HIV / AIDS patients in China are both mutated but still at low level. The amino acid positions of conserved epitopes of different types of neutralizing antibodies The degree of variation is different.